Temporal distribution of the ganglion cell volleys in the normal rat optic nerve

We describe experiments on behaving rats with electrodes implanted on the cornea, in the optic chiasm, and on the visual cortex; in addition, two red light-emitting diodes (LED) are permanently attached to the skull over the left eye. Recordings timelocked to the LED flashes reveal both the local events at each electrode site and the orderly transfer of visual information from retina to cortex. The major finding is that every stimulus, regardless of its luminance, duration, or the state of retinal light adaptation, elicits an optic nerve volley with a latency of about 10 ms and a duration of about 300 ms. This phenomenon has not been reported previously, so far as we are aware. We conclude that the retina, which originates from the forebrain of the developing embryo, behaves like a typical brain structure: it translates, within a few hundred milliseconds, the chemical information in each pattern of bleached photoreceptors into a corresponding pattern of ganglion cell neuronal information that leaves via the optic nerve. The attributes of each rat ganglion cell appear to include whether the retinal neuropile calls on it to leave after a stimulus and, if so when, within a 300-ms poststimulus epoch. The resulting retinal analysis of the scene, on arrival at the cortical level, is presumed to participate importantly in the creation of visual perceptual experiences.

Sleep modifies retinal ganglion cell responses in the normal rat

Recordings were obtained from the visual system of rats as they cycled normally between waking (W), slow-wave sleep (SWS), and rapid eye movement (REM) sleep. Responses to flashes delivered by a light-emitting diode attached permanently to the skull were recorded through electrodes implanted on the cornea, in the chiasm, and on the cortex. The chiasm response reveals the temporal order in which the activated ganglion cell population exits the eyeball; as reported, this triphasic event is invariably short in latency (5–10 ms) and around 300 ms in duration, called the histogram. Here we describe the differences in the histograms recorded during W, SWS, and REM. SWS histograms are always larger than W histograms, and an REM histogram can resemble either. In other words, the optic nerve response to a given stimulus is labile; its configuration depends on whether the rat is asleep or awake. We link this physiological information with the anatomical fact that the brain dorsal raphe region, which is known to have a sleep regulatory role, sends fibers to the rat retina and receives fibers from it. At the cortical electrode, the visual cortical response amplitudes also vary, being largest during SWS. This well known phenomenon often is explained by changes taking place at the thalamic level. However, in the rat, the labile cortical response covaries with the labile optic nerve response, which suggests the cortical response enhancement during SWS is determined more by what happens in the retina than by what happens in the thalamus.

p63 identifies keratinocyte stem cells

The proliferative compartment of stratified squamous epithelia consists of stem and transient amplifying (TA) keratinocytes. Some polypeptides are more abundant in putative epidermal stem cells than in TA cells, but no polypeptide confined to the stem cells has yet been identified. Here we show that the p63 transcription factor, a p53 homologue essential for regenerative proliferation in epithelial development, distinguishes human keratinocyte stem cells from their TA progeny. Within the cornea, nuclear p63 is expressed by the basal cells of the limbal epithelium, but not by TA cells covering the corneal surface. Human keratinocyte stem and TA cells when isolated in culture give rise to holoclones and paraclones, respectively. We show by clonal analysis that p63 is abundantly expressed by epidermal and limbal holoclones, but is undetectable in paraclones. TA keratinocytes, immediately after their withdrawal from the stem cell compartment (meroclones), have greatly reduced p63, even though they possess very appreciable proliferative capacity. Clonal evolution (i.e., generation of TA cells from precursor stem cells) is promoted by the sigma isoform of the 14-3-3 family of proteins. Keratinocytes whose 14-3-3σ has been down-regulated remain in the stem cell compartment and maintain p63 during serial cultivation. The identification of p63 as a keratinocyte stem cell marker will be of practical importance for the clinical application of epithelial cultures in cell therapy as well as for studies on epithelial tumorigenesis.

Corneal collagen fibril structure in three dimensions: Structural insights into fibril assembly, mechanical properties, and tissue organization

The ability of the cornea to transmit light while being mechanically resilient is directly attributable to the formation of an extracellular matrix containing orthogonal sheets of collagen fibrils. The detailed structure of the fibrils and how this structure underpins the mechanical properties and organization of the cornea is understood poorly. In this study, we used automated electron tomography to study the three-dimensional organization of molecules in corneal collagen fibrils. The reconstructions show that the collagen molecules in the 36-nm diameter collagen fibrils are organized into microfibrils (≈4-nm diameter) that are tilted by ≈15° to the fibril long axis in a right-handed helix. An unexpected finding was that the microfibrils exhibit a constant-tilt angle independent of radial position within the fibril. This feature suggests that microfibrils in concentric layers are not always parallel to each other and cannot retain the same neighbors between layers. Analysis of the lateral structure shows that the microfibrils exhibit regions of order and disorder within the 67-nm axial repeat of collagen fibrils. Furthermore, the microfibrils are ordered at three specific regions of the axial repeat of collagen fibrils that correspond to the N- and C-telopeptides and the d-band of the gap zone. The reconstructions also show macromolecules binding to the fibril surface at sites that correspond precisely to where the microfibrils are most orderly.

Influencia de las medidas queratométricas en el cálculo de la potencia de lentes intraoculares

Índice, resumen, conclusiones y bibliografía de la memoria del Máster en Optometría Clínica y Visión, Programa formativo en Biomedicina y Tecnologías para la vida.

Corneal topography reinterpretation through separate analysis of the projected rings

Póster presentado en SPIE Photonics Europe, Brussels, 16-19 April 2012.

In vivo characterization of corneal biomechanics

Interest in corneal biomechanics has increased with the development of new refractive surgery techniques aimed at modifying corneal properties and a variety of surgical options for corneal ectasia management. The human cornea behaves as soft biological material. It is a viscoelastic tissue and its response to a force applied to it depends not only on the magnitude of the force, but also on the velocity of the application. There are concerns about the limitations to measuring corneal biomechanical properties in vivo. To date, 2 systems are available for clinical use: the Ocular Response Analyzer, a dynamic bidirectional applanation device, and the Corvis ST, a dynamic Scheimpflug analyzer device. These devices are useful in clinical practice, especially for planning some surgical procedures and earlier detection of ectatic conditions, but further research is needed to connect the clinical measurements obtained with these devices to the standard mechanical properties.

Screening Subclinical Keratoconus With Placido-Based Corneal Indices

Purpose. To assess in a sample of normal, keratoconic, and keratoconus (KC) suspect eyes the performance of a set of new topographic indices computed directly from the digitized images of the Placido rings. Methods. This comparative study was composed of a total of 124 eyes of 106 patients from the ophthalmic clinics Vissum Alicante and Vissum Almería (Spain) divided into three groups: control group (50 eyes), KC group (50 eyes), and KC suspect group (24 eyes). In all cases, a comprehensive examination was performed, including the corneal topography with a Placidobased CSO topography system. Clinical outcomes were compared among groups, along with the discriminating performance of the proposed irregularity indices. Results. Significant differences at level 0.05 were found on the values of the indices among groups by means of Mann-Whitney-Wilcoxon nonparametric test and Fisher exact test. Additional statistical methods, such as receiver operating characteristic analysis and K-fold cross validation, confirmed the capability of the indices to discriminate between the three groups. Conclusions. Direct analysis of the digitized images of the Placido mires projected on the cornea is a valid and effective tool for detection of corneal irregularities. Although based only on the data from the anterior surface of the cornea, the new indices performed well even when applied to the KC suspect eyes. They have the advantage of simplicity of calculation combined with high sensitivity in corneal irregularity detection and thus can be used as supplementary criteria for diagnosing and grading KC that can be added to the current keratometric classifications.

Reverse Geometry Hybrid Contact Lens Fitting in a Case of Donor-Host Misalignment after Keratoplasty

Purpose: To report the successful outcome obtained after fitting a new hybrid contact lens in a cornea with an area of donor-host misalignment and significant levels of irregular astigmatism after penetrating keratoplasty (PKP). Materials and methods: A 41-year-old female with bilateral asymmetric keratoconus underwent PKP in her left eye due to the advanced status of the disease. One year after surgery, the patient referred a poor visual acuity and quality in this eye. The fitting of different types of rigid gas permeable contact lenses was performed, but with an unsuccessful outcome due to contact lens stability problems and uncomfortable wear. Scheimpflug imaging evaluation revealed that a donor-host misalignment was present at the nasal area. Contact lens fitting with a reverse geometry hybrid contact lens (Clearkone, SynergEyes Carlsbad) was then fitted. Visual, refractive, and ocular aberrometric outcomes were evaluated during a 1-year period after the fitting. Results: Uncorrected distance visual acuity improved from a prefitting value of 20/200 to a best corrected postfitting value of 20/20. Prefitting manifest refraction was +5.00 sphere and -5.50 cylinder at 75°, with a corrected distance visual acuity of 20/30. Higher order root mean square (RMS) for a 5 mm pupil changed from a prefitting value of 6.83 µm to a postfitting value of 1.57 µm (5 mm pupil). The contact lens wearing was referred as comfortable, with no anterior segment alterations. Conclusion: The SynergEyes Clearkone contact lens seems to be another potentially useful option for the visual rehabilitation after PKP, especially in cases of donor-host misalignment.

Technologies for Anatomical and Geometric Characterization of the Corneal Structure and Anterior Segment: A Review

Corneal and anterior segment imaging techniques have become a crucial tool in the clinical practice of ophthalmology, with a great variety of applications, such as corneal curvature and pachymetric analysis, detection of ectatic corneal conditions, anatomical study of the anterior segment prior to phakic intraocular lens implantation, or densitometric analysis of the crystalline lens. From the Placido-based systems that allow only a characterization of the geometry of the anterior corneal surface to the Scheimpflug photography-based systems that provide a characterization of the cornea, anterior chamber, and crystalline lens, there is a great variety of devices with the capability of analyzing different anatomical parameters with very high precision. To date, Scheimpflug photography-based systems are the devices providing the more complete analysis of the anterior segment in a non-invasive way. More developments are required in anterior segment imaging technologies in order to improve the analysis of the crystalline lens structure as well as the ocular structures behind the iris in a non-invasive way when the pupil is not dilated.

Repeatability and reproducibility of corneal thickness using SOCT Copernicus HR

Purpose: The aim of this study is to determine the reliability of corneal thickness measurements derived from SOCT Copernicus HR (Fourier domain OCT). Methods: Thirty healthy eyes of 30 subjects were evaluated. One eye of each patient was chosen randomly. Images were obtained of the central (up to 2.0 mm from the corneal apex) and paracentral (2.0 to 4.0 mm) cornea. We assessed corneal thickness (central and paracentral) and epithelium thickness. The intra-observer repeatability data were analysed using the intra-class correlation coefficient (ICC) for a range of 95 per cent within-subject standard deviation (SW) and the within-subject coefficient of variation (CW). The level of agreement by Bland–Altman analysis was also represented for the study of the reproducibility between observers and agreement between methods of measurement (automatic versus manual). Results: The mean value of the central corneal thickness (CCT) was 542.4 ± 30.1 μm (SD). There was a high intra-observer agreement, finding the best result in the central sector with an intra-class correlation coefficient of 0.99, 95 per cent CI (0.989 to 0.997) and the worst, in the minimum corneal thickness, with an intra-class correlation coefficient of 0.672, 95 per cent CI (0.417 to 0.829). Reproducibility between observers was very high. The best result was found in the central sector thickness obtained both manually and automatically with an intra-class correlation coefficient of 0.990 in both cases and the worst result in the maximum corneal thickness with an intra-class correlation coefficient of 0.827. The agreement between measurement methods was also very high with intra-class correlation coefficient greater than 0.91. On the other hand the repeatability and reproducibility for epithelial measurements was poor. Conclusion: Pachymetric mapping with SOCT Copernicus HR was found to be highly repeatable and reproducible. We found that the device lacks an appropriate ergonomic design as proper focusing of the laser beam onto the cornea for anterior segment scanning required that patients were positioned slightly farther away from the machine head-rest than in the setup for retinal imaging.

Algorithm for Correcting the Keratometric Error in the Estimation of the Corneal Power in Eyes With Previous Myopic Laser Refractive Surgery

Purpose: To calculate theoretically the errors in the estimation of corneal power when using the keratometric index (nk) in eyes that underwent laser refractive surgery for the correction of myopia and to define and validate clinically an algorithm for minimizing such errors. Methods: Differences between corneal power estimation by using the classical nk and by using the Gaussian equation in eyes that underwent laser myopic refractive surgery were simulated and evaluated theoretically. Additionally, an adjusted keratometric index (nkadj) model dependent on r1c was developed for minimizing these differences. The model was validated clinically by retrospectively using the data from 32 myopic eyes [range, −1.00 to −6.00 diopters (D)] that had undergone laser in situ keratomileusis using a solid-state laser platform. The agreement between Gaussian (PGaussc) and adjusted keratometric (Pkadj) corneal powers in such eyes was evaluated. Results: It was found that overestimations of corneal power up to 3.5 D were possible for nk = 1.3375 according to our simulations. The nk value to avoid the keratometric error ranged between 1.2984 and 1.3297. The following nkadj models were obtained: nkadj= −0.0064286r1c + 1.37688 (Gullstrand eye model) and nkadj = −0.0063804r1c + 1.37806 (Le Grand). The mean difference between Pkadj and PGaussc was 0.00 D, with limits of agreement of −0.45 and +0.46 D. This difference correlated significantly with the posterior corneal radius (r = −0.94, P < 0.01). Conclusions: The use of a single nk for estimating the corneal power in eyes that underwent a laser myopic refractive surgery can lead to significant errors. These errors can be minimized by using a variable nk dependent on r1c.

Correlation of the Corneal Toricity Between Anterior and Posterior Corneal Surfaces in the Normal Human Eye

Purpose: To evaluate the correlation of the magnitude of corneal toricity and power vector components of both corneal surfaces measured with a Scheimpflug photography-based system. Methods: A total of 117 healthy normal eyes of 117 subjects selected randomly with ages ranging from 7 to 80 years were included. All eyes received an anterior segment and corneal analysis with the Sirius system (CSO) evaluating the anterior and posterior mean toricity for 3 and 7 mm (aAST and pAST). The vector components J0 and J45 as well as the overall strength blur (B) were calculated for each keratometric measurement using the procedure defined by Thibos and Horner. Results: The coefficient of correlation between aAST and pAST was 0.52 and 0.62 and the mean anteroposterior ratio for toricity was 0.46 ± 0.39 and 0.57 ± 0.75 for 3 and 7 mm, respectively. These ratios correlated significantly with aAST, anterior corneal J0, and manifest refraction J0 (r ≥ 0.39, P < 0.01). The coefficient of correlation was 0.69 and 0.81 between anterior and posterior J0 for 3 and 7 mm, respectively. For J45, the coefficients were 0.62 and 0.71, respectively. The linear regression analysis revealed that the pAST and power vectors could be predicted from the anterior corneal data (R2 ≥ 0.40, P < 0.01). Conclusions: The toricity and astigmatic power vector components of the posterior corneal surface in the human healthy eye are related to those of the anterior and therefore can be predicted consistently from the anterior toricity and astigmatic power vectors.

Correlation of Anterior and Posterior Corneal Shape in Keratoconus

Purpose: To evaluate the correlation of the mean curvature and shape factors of both corneal surfaces for different corneal diameters measured with the Scheimpflug photography–based system in keratoconus eyes. Methods: A total of 61 keratoconus eyes of 61 subjects, aged 14 to 64 years, were included in this study. All eyes received a comprehensive ophthalmologic examination including anterior segment and corneal analysis with the Sirius system (CSO): anterior and posterior mean corneal radius for 3, 5, and 7 mm (aKM, pKM), anterior and posterior mean shape factor for 4.5 and 8 mm (ap, pp), central and minimal corneal thickness, and anterior chamber depth. Results: Mean aKM/pKM ratio around 1.20 (range, 0.95–1.48) was found for all corneal diameters (P = 0.24). Weak but significant correlations of this ratio with pachymetric parameters were found (r between −0.28 and −0.34, P < 0.04). The correlation coefficient between aKM and pKM was ≥0.92 for all corneal diameters. A strong and significant correlation was also found between ap and pp (r ≥ 0.86, P < 0.01). The multiple regression analysis revealed that central pKM was significantly correlated with aKM, central corneal thickness, anterior chamber depth, and spherical equivalent (R2 ≥ 0.88, P < 0.01) and that 8 mm pp was significantly correlated with 8 mm ap and age (R2 = 0.89, P < 0.01). Conclusions: Central posterior corneal curvature and shape factor in the keratoconus eye can be consistently predicted from the anterior corneal curvature and shape factor, respectively, in combination with other anatomical and ocular parameters.

First EURONEAR NEA discoveries from La Palma using the INT

Since 2006, the European Near Earth Asteroids Research (EURONEAR) project has been contributing to the research of near-Earth asteroids (NEAs) within a European network. One of the main aims is the amelioration of the orbits of NEAs, and starting in 2014 February we focus on the recovery of one-opposition NEAs using the Isaac Newton Telescope (INT) in La Palma in override mode. Part of this NEA recovery project, since 2014 June EURONEAR serendipitously started to discover and secure the first NEAs from La Palma and using the INT, thanks to the teamwork including amateurs and students who promptly reduce the data, report discoveries and secure new objects recovered with the INT and few other telescopes from the EURONEAR network. Five NEAs were discovered with the INT, including 2014 LU14, 2014 NL52 (one very fast rotator), 2014 OL339 (the fourth known Earth quasi-satellite), 2014 SG143 (a quite large NEA), and 2014 VP. Another very fast moving NEA was discovered but was unfortunately lost due to lack of follow-up time. Additionally, another 14 NEA candidates were identified based on two models, all being rapidly followed-up using the INT and another 11 telescopes within the EURONEAR network. They include one object discovered by Pan-STARRS, two Mars crossers, two Hungarias, one Jupiter trojan, and other few inner main belt asteroids (MBAs). Using the INT and Sierra Nevada 1.5 m for photometry, then the Gran Telescopio de Canarias for spectroscopy, we derived the very rapid rotation of 2014 NL52, then its albedo, magnitude, size, and its spectral class. Based on the total sky coverage in dark conditions, we evaluate the actual survey discovery rate using 2-m class telescopes. One NEA is possible to be discovered randomly within minimum 2.8 deg2 and maximum 5.5 deg2. These findings update our past statistics, being based on double sky coverage and taking into account the recent increase in discovery.